Training Guide for NG-Test® CARBA 5

January 04, 2024

Posted: January 04, 2024

Categories: Laboratory Tutorials, Modern Innovations, Featured Posts, Clinical, Industry Insights, Product Spotlights

Tags: tutorial, training guide, ng-test, cre, carba5, carba 5, amr

Carbapenem-resistant Enterobacterales or CRE, are a group of gram-negative bacteria that are resistant to “last line” antibiotics and are very challenging to treat. These microorganisms may confer their resistance via the production of a carbapenemase, an enzyme that inactivates carbapenem antibiotics.

The five most common types of carbapenemases are KPC, OXA-48-like, VIM, IMP, and NDM.

Knowing the type of carbapenemase is crucial for therapeutic purposes,
infection control, and for preventing outbreaks.

Different enzymes require different antibiotic regimens and some enzymes can more easily spread through hospitals or networks of hospitals, leading to an epidemic spread of antimicrobial resistance genes.

CRE infections have an estimated mortality rate of up to 50%, therefore it is imperative to know if and what type of carbapenemase is being produced by a pathogen, quickly and accurately.

Hardy Diagnostics is the exclusive distributor in the United States of the rapid, multiplex lateral flow assay, the NG-Test® CARBA 5. In just 15 minutes, CARBA 5 is able to detect and differentiate the “big five” carbapenemases produced by Enterobacterales and Pseudomonas aeruginosa.

This FDA cleared device delivers phenotypic, (not genotypic), test results which is more relevant to the disease status, in just 15 minutes using colonies from HardyCHROM™ CRE agar, Blood agar or MacConkey agar.

The Advantages of NG-Test® CARBA 5

Rapid

With about one minute of hands-on time and results in 15 minutes per test, NG-Test® CARBA 5 naturally fits into any laboratory workflow.

Accurate

Unlike molecular methods,NG-Test® CARBA 5 detects the carbapenemase enzyme which is crucial to aiding therapeutic decisions.

Additionally,NG-Test® CARBA 5 has been able to detect several variants of carbapenemases, undetected by predicate PCR methods.

User Friendly

This lateral flow assay is incredibly easy to use and requires no specialized training or additional equipment, unlike molecular assays.

Catalog no. CARBA5

Other benefits of NG-Test® CARBA 5:

There are no maintenance costs associated with the device and the product is stable at room temperature with a two-year shelf life.
Each kit of NG-Test® CARBA 5 comes with all the equipment required for use.
NG-Test® CARBA 5 can be used in place of molecular detection methods. Molecular methods require specialized technician training, are very expensive per test, and can take about two to three hours for a result.
Additionally, molecular assays detect the presence of particular genes, but do not indicate whether or not the gene is being expressed.

NG-Test® CARBA 5 detects and differentiates carbapenemases
produced by microorganisms to deliver a phenotypic result.

mCIM and eCIM assays detect and differentiate between the two major groups of carbapenemases: serine beta-lactamases, like KPC and OXA-48-like, and metallo-beta-lactamases, like VIM, IMP, and NDM. However, CIM tests are unable to further differentiate specific enzymes.
Additionally, in the mCIM and eCIM assay, serine beta-lactamases will mask metallo-beta-lactamases if the enzymes are co-produced. The Carba NP test, another phenotypic assay, has similar limitations to CIM test methods in that it does not differentiate the specific carbapenemase that is produced.

NG-Test® CARBA 5 solves this problem by detecting and differentiating
multiple carbapenemases in a single assay.

Protocol Demonstration

STEP 1

Add 5 drops of Extraction Buffer to the tube

STEP 2

Using a 1μl loop, touch 3 colonies and inoculate the tube with buffer.

STEP 3

Vortex until the suspension is homogenous, this is important to ensure that the cells have lysed.

STEP 4

Open the mylar pouch containing the NG-Test CARBA 5 lateral flow device.

STEP 5

Using the supplied transfer pipette, draw the sample until it reaches the 100μl black line.

STEP 6

Dispense the sample into the sample well labeled “S” and start your timer. Allow the sample to diffuse along the test strip. Read the results at 15 minutes.